Determining the Structure, Function and Regulation of Dynein and Flagella

确定动力蛋白和鞭毛的结构、功能和调节

基本信息

批准号：
8529556
负责人：
Daniela Nicastro
金额：
$ 31.05万
依托单位：
BRANDEIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-28 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8529556
关键词：
Address Attenuated Bardet-Biedl Syndrome Behavior Binding Biochemical Biological Budgets Cell division Cell physiology Cells Chlamydomonas Cilia Classification Complex Congenital Disorders Congenital Heart Defects Cytoskeleton Data Defect Development Disease Docking Dynein ATPase Esthesia Flagella Freezing Genes Genetic Goals Head Heterogeneity Homologous Gene Human In Situ Individual Knock-out Knowledge Label Laboratories Lead Length Life Life Cycle Stages Link Lung diseases Malignant Neoplasms Malignant neoplasm of brain Maps Mass Spectrum Analysis Microtubules Molecular Molecular Motors Morphogenesis Motor Movement Neurodegenerative Disorders Organ Organelles Organism Play Polycystic Kidney Diseases Primary Ciliary Dyskinesias Process Proteins Proteome Protocols documentation Radial Regulation Research Role Sea Urchins Signal Pathway Signal Transduction Situs Inversus Slide Sperm Tail Spottings Stretching Structural Biochemistry Structure System Techniques Tetrahymena Triplet Multiple Birth Ursidae Family Work arm base cell motility comparative density electron tomography image processing improved innovation insight molecular imaging mutant nexin protein function therapeutic development three dimensional structure trafficking

项目摘要

DESCRIPTION (provided by applicant): Cilia and flagella are evolutionarily conserved organelles with important biological roles in motility, sensation and signaling. They are essential in the life cycles of most eukaryotic organisms, and cilia are found in nearly every human cell. The goal of this research is to determine how regulatory signals change the activity of the molecular motor protein dynein, and how dynein activity is regulated in motile cilia and flagella to generate proper beating. Our laboratory opened a new window into the three-dimensional organization and protein composition of cilia and flagella, and recently discovered previously unseen structures that regulate dynein function and/or are essential for flagellar assembly. Building on a strong base of preliminary data gathered in the preceding project period, this proposal directly addresses key open questions in the field in three specific aims that are directed at understanding the three-dimensional structure, subunit composition, protein interactions and the regulatory functions of the following three major regulatory and signal transduction complexes in cilia and flagella: (1) the nexin-dynein regulatory complex, (2) the I1 inner dynein complex, and (3) radial spoke 3 in organisms with radial spoke triplets. The project uses a multi-disciplinary approach and cutting-edge techniques, combining in situ molecular imaging by cryo-electron tomography and image processing, biochemical and mass spectrometric analyses, integrated structural-genetics approaches and protein labeling techniques to directly visualize gene products in cells. The proposed research will contribute fundamental knowledge and a deeper understanding of the mechanisms underlying motor protein function and control on a molecular level and of the functional organization of cilia and flagella.

描述（由申请人提供）：纤毛和鞭毛是进化保守的细胞器，在运动，感觉和信号传导中具有重要的生物学作用。它们在大多数真核生物的生命周期中至关重要，几乎每个人类细胞都发现了纤毛。这项研究的目的是确定调节信号如何改变分子运动蛋白动力蛋白的活性，以及如何调节纤毛和鞭毛中的动力蛋白活性以产生适当的打击。我们的实验室为纤毛和鞭毛的三维组织和蛋白质组成打开了一个新窗口，最近发现了先前看不见的结构，这些结构调节了动力蛋白功能和/或对于鞭毛组装至关重要。该提案以在前一个项目期间收集的强大初步数据基础为基础，直接解决了领域中的关键开放问题，这些特定目的是针对理解三维结构，亚基组成，蛋白质相互作用，蛋白质相互作用以及以下三个主要调节和信号传输复合物中的cilia和flagella i Innexinne n neyxine nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexin nexine（1）复合物，（3）径向在带有径向辐条三重的生物体中说3。该项目采用多学科方法和最先学的技术，通过冷冻电子断层扫描和图像处理，生化和质谱分析，集成结构基因方法和蛋白质标记技术来直接可视化细胞中的基因产物，结合原位分子成像。拟议的研究将有助于基本知识，并更深入地了解运动蛋白功能和控制纤毛和鞭毛的功能组织的机制。